Study On The Extraction Technology Of Buyang Huanwu Decoction Based On The Kinetic Model Of Flos Carthami And Radix Paeoniae Rubra

Objective:A decoction kinetic model of Flos Carthami and Radix Paeoniae Rubra in Buyang Huanwu Decoction was established based on Fick rule of diffusionand the use of TCL and HPLC to screen out Flos Carthami and Radix Paeoniae Rubra in the be st place of origin.On this basis,the factors affecting the extraction process of Buyang Huanwu decoction were investigated,and the optimum extraction process was determined by Orthogonal test.Methods:1.Determining the best place of origin of Flos Carthami.Purchased two sets of Flos Carthami from Jianyang City of Sichuan Province,Yumin County of Xinjiang and Yumen City of Gansu Province respectively;and one set of Flos Carthami from Jimsar County of Xinjiang,Yongsheng County of Yunnan,Weishan County of Yunnan Province,and Yining city of Xinjiang respectively.Atotal of 10 sets of Flos Carthami were tested by thin layer chromatography,and the hydroxysafflor yellow A content in the Flos Carthami samples were compared by high performance liquid chromatography.The thin layer conditions were as follow:taking 5?l of the above ten kinds of solutions and place on the same silica gel H thin layer plate at different spots;using ethyl acetate-formic acid-water-methanol(7:2:3:0.4)as a developing solvent.The chromatographic conditions were as follow:column;Agilent C18 column(250mm�4.6mm,5?m);mobile phase was methanol-acetonitrile-0.7%phosphoric acid solution(26:2:72);flow rate:lml/min;detection wavelength:403nm;column Temperature:25?;injection volume:10?l.2.Determining the best place of origin of Radix Paeoniae Rubra.Purchased one set of Radix Paeoniae Rubra from Helen County of Inner Mongolia,Xifeng County of Liaoning Province,Siping County of Jilin Province,Zhangzhou of Anhui Province,Daqing of Heilongjiang Province,Olunchun Banner of Inner Mongolia,Fengcheng of Liaoning Province,Jiamusi City of Heilongjiang Province,Aershan of Inner Mongolia,and Xing'an League of Inner Mongolia Akasaka respectively.A total of 10 sets of Radix Paeoniae Rubra were analyzed by thin layer chro matography.The paenoiflorin contained in the mixture were compared by high perfo rmance liquid chromatograph.The thin layer conditions were as follow:taking 4?l of each of the above ten kinds of solutions,and placing on the same silica gel G thin layer plate at different spots;using chloroform-ethyl acetate-methanol-formic acid(40:5:10:0.2)as a developing solvent.To unfold,remove,dry,and spray it with 5%vanillin sulfuric acid solution and to heat until the spots are clear.The chromatogr aphic conditions were as follow:column:Agilent C18 column(250mm�4.6mm,5?m);mobile phase was methanol-0.05 mol/L potassium dihydrogen phosphate solution(40:65);flowrate:lml/min;detection wavelength:230nm;Column temperature:30?;injection volume:10?l.3.To establish a method for the determination of four active components in Buyang Huanwu decoction.Taking hydroxysafflor yellow A,flavonoid glucoside,paeoniflorin and ferulic acid as four indicators for determining the content of components in the liquid,establishing HPCL assay for Buyang Huanwu Decoction column.4.Establishment of decoction kinetic model of Flos Carthami membranaceus in Buyang Huanwu DecoctionThe decoction kinetics of hydroxysafflor yellow A,the main active ingredient of Flos Carthami sinensis,was studied in Buyang Huanwu Decoction.The decoction kinetics model was established.Based on Fick rule of diffusion,the equation parameters were derived and the measurement results were substituted into the mathematic al model (?),and equation fitting was performed.5.Establishment of decoction kinetic model of Paeoniae Rubra membranaceus in Buyang Huanwu DecoctionThe decoction kinetics of ferulic acid,the main active ingredient of Paeoniae Rubra,was studied in Buyang Huanwu Decoction.The decoction kinetics model was established.Based on Fick rule of diffusion,the equation parameters were derived and the measurement results were substituted into the mathematical model (?),and equation fitting was performed.6.Optimization of the best extraction process of Buyang Huanwu DecoctionHydroxysafflor yellow A,flavonoid glucoside,paeoniflorin and ferulic acid were taken as four indicators to determent the content of components in the liquid,Under the condition of liquid phase,The orthogonal test was carried out by orthogonal test method according to the L9(34)orthogonal table for the multiple amount of solvent,extraction time and extraction times.With the orthogonal test and decoction keneic model,the best extraction process was then determined.7.The data in this paper was analyzed by using Microsoft Office Excel 2016 and SPSS 17.0 software.The statistical methods were used for the analysis of measurement data,linear regression analysis and orthogonal test design.Results:1.Determining the best place of origin of Flos CarthamiThe thin layer results of ten medicinal materials in the corresponding positions are all showing spots with same color.The highest content is from sample "Jianyang City of Sichuan Province 2",which contains hydroxysafflor yellow A 2.58%.Based on the results of this experiment,the content of hydroxysafflor yellow A in safflower medicinal materials was ranked from the highest content to the lowest as the following:Jianyang City of Sichuan Province 2>Jianyang City of Sichuan Province 1>Yumin County of Xinjiang 1>Yumin County of Xinjiang 2>Jimsar County of Xinjiang>Yongsheng County of Yunnan Province>Weishan County of Yunnan Province>Yumen City of Gansu Province 2>Yumen City of Gansu Province 1>Yining city of Xinjiang.2.Determining the best place of origin of Radix Paeoniae RubraThe thin layer results of ten medicinal materials in the corresponding positions,all showing spots with same color.The highest content was from sample "Duolun County of Inner Mongolia";the content of paeoniflorin was 4.07%.According to the results of this experiment,the contents of paeoniflorin in Radix Paeoniae Alba were ranked from the highest content to the lowest as the following:Duolun County f Inner Mongolia>Xifeng County of Liaoning Province>Siping County of Jilin Province>Cangzhou of Anhui Province>Daqing of Heilongjiang Province>Europe an Banner of Inner Mongolia>Fengcheng of Liaoning Province>Jiamusi City of Heilongjiang Province>Aershan of Inner Mongolia>Xing'an League of Inner Mongolia.3.To establish a method for the determination of four active components in Buyang Huanwu decoction.HPLC method was used for the determination of hydroxysafflor yellow a,Glucoside,ferulic acid and Paeoniflorin in Buyang Huanwu decoction.The chromatographic conditions were as follows:Agilent Technologies C18 column Nu-Analytical,250 mm�4.6 mm�5 m;Mobile Phase:Acetonitrile(A)-0.05%phosphoric acid aqueous solution(B),gradient elution(0?30min,5?20%A,95?80%B;30?60min,20?60%A,80?40%B;60?70min,60?90%A,40?10%B;70?80min,90%A,10%B);flow rate:1 ml/Min;detection wavelength:280 Nm;injection volume:10 ?l.The method has good precision,stability and repeatability and is feasible in operation.4.Establishment of decoction kinetic model of Flos Carthami membranaceus in Buyang Huanwu DecoctionThe equation is calculated as follows:(?),the verification results can well describe the dynamic process of Flos Carthami in buyanghuanwu decoction,and the standard deviation between the test value and the calculated value is 4.37%,which can be controlled within the required range of industrial standard deviation less than 10%.5.Establishment of decoction kinetic model of Radix Paeoniae Rubra membranaceus in Buyang Huanwu DecoctionThe equation is deduced as follows:(?),the result can describe the kinetic process of Radix paeoniae Rubra in Buyang Huanwu decoction well,the standard deviation between the experimental value and the calculated value is 0.46%,and can be controlled in the industrial standard deviation less than 10%.6.The optimum extraction process of Buyang Huanwu decoction was optimizedThe SPSS software was used to analyze the test data.The effects of three factors on the extraction of hydroxysafflor yellow A from the highest to the lowest were C>B>A.The effects of three factors on the extraction of glucosinolates from the highest to the lowest were B>C>A.The effects of three factors on the extraction of paeoniflorin from the highest to the lowest were A>B>C.The influence of three factors on.the extraction of ferulic acid from the highest to the lowest were C>A>B.As a result,the optimal extraction process is A2B1C2,which is 8 times of the amount of water.The solvent was extracted 60 minutes each time,and extracted twice.Conclusion:The best place of origin of Flos Carthami and the best place of origin of Paeoniae Rubra is determined,which provided the basis and quality assurance for the study of Buyang Huanwu Decoction.The successful establishment of the decoction dynamics model of Flos Carthami and Radix Paeoniae Rubra in Buyang Huanwu Decoction,which can provide experimental basis and basis for further research on the extraction process of Buyang Huanwu Decoction.